Abstract
In this tutorial mini-review, we explore the application of Design of Experiments (DOE) as a powerful statistical tool in biotechnology. Specifically, we review the optimization of hydrogel materials for diverse microbial applications related to green microbiology, the use of microbes to promote sustainability. Hydrogels, three-dimensional polymers networks with high water retention capabilities, are pivotal in the immobilization of microorganisms and provide a customizable environment essential for directing microbial fate. We focus on the application of DOE to precisely tailor hydrogel compositions for a range of fungi and bacteria either used for the sustainable production of chemical compounds, or the elimination of hazardous substances. We examine a variety of DOE design strategies such as central composite designs, Box-Behnken designs, and optimal designs, and discuss their strategic implementation across diverse hydrogel formulations. Our analysis explores the integral role of DOE in refining hydrogels derived from a spectrum of polymers, including natural and synthetic polymers. We illustrate how DOE facilitates nuanced control over hydrogel properties that cannot be achieved using a standard one factor at a time approach. Furthermore, this review reveals a conserved finding across different materials and applications: there are significant interactions between hydrogel parameters and cell behavior. This highlights the intricacies of cell-hydrogel interactions and the impact on hydrogel material properties and cellular functions. Lastly, this review not only highlights DOE's efficacy in streamlining the optimization of cell-hydrogel processes but also positions it as a critical tool in advancing our understanding of cell-hydrogel dynamics, potentially leading to innovative advancements in biotechnological applications and bioengineering solutions.